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AP_Bio_-_Spectrophotometry

AP_Bio_-_Spectrophotometry - Applications of...

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Spectrophotometry
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Does a spectrophotomer do? Detects how much light passes through a sample Who cares about this? Gives you insight into what is in your sample
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How does this work?
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Vary the wavelength (keep conc of sample the same) Determine the “absorption spectrum” Like a molecular fingerprint – λ max is easy to spot – place where absorbance is the highest % Transmittance can be converted to absorption
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Vary the concentration (keep wavelength the same) Absorption is directly proportional to concentration
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Beer’s law relates absorption to concentraiton A = ε c l ε – molar absorptivity c – concentration of sample l – path length for sample
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Unformatted text preview: Applications of spectrophotometry are varied: • Some examples – Determine the absorption spectrum for a newly discovered molecule – Food & drug analysis • WHAT is in a sample – Measuring concentration of a sample in solution – Measuring how the concentration of a sample changes over time • Ex – measure change in concentration of a substrate or product to determine reaction rate Consider…. . • What limitations does this have for us when studying an enzyme mediated reaction? • Substance must – Be Colored – Be Dissolved in solution – Have a λ max that does not coincide with other molecules in the solution...
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AP_Bio_-_Spectrophotometry - Applications of...

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